WO2003042187A1

WO2003042187A1 - Serin protease inhibitors

Info

Publication number: WO2003042187A1
Application number: PCT/EP2002/012697
Authority: WO
Inventors: Katrin Illgen; Thilo Fuchs; Sven Nerdinger
Original assignee: Morphochem Aktiengesellschaft für kombinatorische Chemie
Priority date: 2001-11-13
Filing date: 2002-11-13
Publication date: 2003-05-22
Also published as: DE10155727A1

Abstract

The invention relates to compounds of formula (I) or a pharmaceutically acceptable salt, solvate, hydrate or a pharmaceutically acceptable formulation thereof. Said compounds can be used to inhibit serin proteases, especially tryptase and uPA, and to prevent and/or treat tryptase and uPA related diseases.

Description

Serine protease inhibitors

The present invention describes new compounds, their pro-drugs, pharmacologically acceptable salts and drug compositions containing the same as an active ingredient. These new compounds are serine protease inhibitors, in particular strong tryptase and uPA inhibitors, which they in the prevention and treatment of diseases in which tryptase is involved such. B. asthma, rheumatoid arthritis, psoriasis or uPA is involved such as in the case of tumor diseases (both local tumors and in the advanced stage) and metastasis.

Tryptases are a family of homologous serine proteases that are particularly abundant in mast cells in a tetrameric complex with sulfated carbohydrates, e.g. B. heparin occur. When mast cells are activated, catalytically active tryptase is released from the mast cells into extracellular liquids.

A number of diseases and disease states are related to the proteolytic activity of tryptase, which is related to the activation of a number of other proteins such as cytokines and enzymes which are then involved in such diseases. Therefore, the new compounds of the present invention, which are tryptase inhibitors, are useful in the treatment and prevention of a number of other diseases by using either alone or in combination with other therapeutically useful agents become. These diseases include or may include: inflammatory diseases of the lung system such as asthma, allergic rhinitis, chronic obstructive pulmonary disease, emphysema, viral and bacterial lung infections and inflammatory reactions (Kyle C. Elrod, Robert P. Numerof, Emerging Therapeutic Targets, 1999, 203-212 ). Other diseases in which tryptase inhibitors can be of therapeutic use are rheumatoid arthritis, psoriasis, inflammatory bowel diseases, multiple sclerosis and cancer. Tryptase is often found in high concentrations in a number of biological fluids and has a relatively long half-life.

The urokinase plasminogen activator (uPA) plays a central role in tumor progression, tumor invasion and metastasis. Malignant tumor cells can detach from a primary tumor and migrate to neighboring tissues, blood vessels or lymph. They are then transported to more distant places, where they cause new tumors (secondary tumor, metastasis, daughter tumor). These secondary tumors ultimately lead to the death of the patient. After formation of the zymogen pro-uPA on the cell surface-bound uPA receptor (uPAR, CD87), uPA is quickly activated by cathepsins B or L. The binding to the receptor focuses the proteolytic activity on the cell surface. Plasminogen is activated by the uPA-uPAR complex to plasmin, which can break down important components of the extracellular matrix. Plasmin is also able to reactivate uPA thanks to a positive feedback mechanism. Furthermore, uPA also activates matrix metalloproteases, which ultimately contribute to the breakdown of basement membrane collagen. These proteolytic processes allow cancer cells to invade neighboring tissues and metastasize. It could also be shown that in a predominant number of tumor types the primary components of the uPA system, uPA, PAI-1 and uPAR are formed by different cell types. Because of the central role of the urokinase-type plasminogen activator (uPA) as well as its receptor (uPAR) and its inhibitor (PAI-1) in these processes, the inhibition of the proteolytic activity of uPA is a promising target in tumor therapy.

Another aspect of the present invention is to provide a new process for the preparation of new piperazine derivatives which, if possible, takes place in the context of a multicomponent reaction and is widely applicable. Multicomponent reactions are of great interest in the pharmaceutical industry, for example, for the production of substance libraries to find lead structures (A. Dömling, I. Ugi, Angew. Chem. 2000, 112, 3300-3344).

The present invention describes compounds, their prodrugs, salts and formulations which are new and have high activity and selectivity.

The present invention comprises compounds of the general formula (I):

wherein

R ^{1 is} an alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aralkyl or heteroaralkyl group, especially heteroalkylarylalkyl;

R ^{2 is} a hydrogen atom, an alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aralkyl or heteroaralkyl group;

R ^{3 is} a hydrogen atom, an alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aralkyl or heteroaralkyl group; and

R ^{4 is} a hydrogen atom, an alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aralkyl or heteroaralkyl group;

or a pharmacologically acceptable salt, solvate, hydrate or a pharmacologically acceptable formulation thereof. The term alkyl refers to a saturated, straight-chain or branched hydrocarbon group which has 1 to 20 carbon atoms, preferably 1 to 12 carbon atoms, particularly preferably 1 to 6 carbon atoms, for example the methyl, ethyl, propyl, isopropyl, isobutyl , tert-butyl, n-hexyl, 2,2-dimethylbutyl or n-octyl group.

The terms alkenyl and alkynyl refer to at least partially unsaturated, straight-chain or branched hydrocarbon groups which have 2 to 20 carbon atoms, preferably 2 to 12 carbon atoms, particularly preferably 2 to 6 carbon atoms, e.g. B. the ethenyl, allyl, acetylenyl, propargyl, isoprenyl or hex-2-enyl group.

The term heteroalkyl refers to an alkyl, an alkenyl or an alkynyl group in which one or more (preferably 1, 2 or 3) carbon atoms or CH or CH ₂ groups are replaced by an oxygen, nitrogen, Phosphorus or sulfur atoms are replaced (preferably oxygen or nitrogen), for example an alkyloxy group such as methoxy or ethoxy, or a methoxymethyl, nitrile, amino, methylcarboxyalkyl ester, carboxyalkyl ester or 2,3-dioxyethyl group , The term heteroalkyl also refers to a carboxylic acid or a group derived from a carboxylic acid, such as. B. acyl, acyloxy, carboxyalkyl, carboxyalkyl esters, for example methyl carboxyalkyl ester, carboxyalkylamide, alkoxycarbonyl or alkoxycarbonyloxy. The term cycloalkyl or cyclo refers to a saturated or partially unsaturated cyclic group which has one or more rings which have 3 to 14 ring carbon atoms, preferably 3 to 10 ring carbon atoms, for example the cyclopropyl or cyclohexyl -, tetralin or cyclohex-2-enyl group.

The term heterocycloalkyl or heterocyclo refers to a cycloalkyl group as defined above, in which one or more (preferably 1, 2 or 3) ring carbon atoms or ring CH or CH ₂ groups are replaced by an oxygen, nitrogen or , Phosphorus or sulfur atom are replaced, and can represent, for example, the piperidine, morpholine, N-methylpiperazine or N-phenylpiperazine group.

The terms alkylcycloalkyl or heteroalkylcycloalkyl refer to groups which, in accordance with the above definitions, contain both cycloalkyl or heterocycloalkyl and also alkyl, alkenyl, alkynyl and / or heteroalkyl groups.

Examples of such groups are alkylcycloalkyl, alkenylcycloalkyl, alkynylcycloalkyl, alkylheterocycloalkyl, alkenylheterocycloalkyl, alkynylheterocycloalkyl, heteroalkylcycloalkyl, heteroalkenylcycloalkyl, heteroalkynylcycloalkyl, heteroalkylheterocycloalkyl, heteroalkoalkyl or heterocycloalkyl, heteroalkoalkyl, heteroalkylcyclo, heteroalkoalkyl, heteroalkylcyclo, heteroalkoalkyl, heteroalkyl or heteroalkyl, heteroalkyl or heterocycloalkyl The term aryl or Ar refers to an aromatic group which has one or more rings which contain 5 or 6 to 14 ring carbon atoms, preferably 5 or 6 to 10 ring carbon atoms, for example a phenyl, naphthyl, 2- , 3- or 4-methoxyphenyl, 2-, 3- or 4-ethoxyphenyl, 4-carboxyphenylalkyl or 4-hydroxyphenyl group.

The term heteroaryl refers to an aryl group in which one or more (preferably 1, 2 or 3) ring carbon atoms or ring CH or CH ₂ groups are replaced by an oxygen, nitrogen, phosphorus or sulfur atom are, for example the 4-pyridyl, 2-imidazolyl, 3-pyrazolyl and isoquinolinyl group.

The terms aralkyl or heteroaralkyl refer to groups which, according to the above definitions, contain both aryl or heteroaryl and also alkyl, alkenyl, alkynyl and / or heteroalkyl and / or cycloalkyl and / or heterocycloalkyl groups , Examples are arylalkyl, arylalkenyl, arylalkynyl, arylheteroalkyl, arylheteroalkenyl, arylheteroalkynyl, heteroarylheteroalkyl, heteroarylheteroalkenyl, heteroarylheteroalkynyl, arylcycloalkyl, heteroaryocycloalkyl, heteroaryocycloalkyl, heteroaryoaryocycloalkylo - alkenyl, heteroarylcycloalkenyl, arylcycloalkynyl, heteroarylcycloalkynyl, arylheteroalkenyl, heteroarylheteroalkenyl, arylheteroalkynyl, heteroarylheteroalkynyl, heteroarylalkyl, heteroalkenyl group and heteroalkenylalkynyl and heteroalkenyl group and the heteroalkenyl group and the heteroalkenyl group saturated or single, double or triple unsaturated. Further examples are, for example, the tetrahydroisoquinolinyl, benzyl, 2- or 3-ethylindolyl or 4-methylpyridino group.

The terms alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl aryl, heteroaryl, aralkyl, heteroalkylarylalkyl and heteroaralkyl also refer to groups in which one or more hydrogen atoms of such groups are represented by fluorine, chlorine or bromine or iodine atoms or OH, SH, NH ₂ , = NH or N0 ₂ groups are replaced. These terms also refer to groups substituted with unsubstituted alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl or heteroaralkyl groups.

Compounds of formula (I) may contain one or more centers of chirality due to their substitution. The present invention therefore encompasses all pure enantiomers and all pure diastereomers, as well as their mixtures in any mixing ratio.

Compounds of the general formula (I) in which R ^{2 is} a hydrogen atom are preferred.

Compounds of the general formula are further preferred

(I), wherein R ^{4 is} a group of the formula -C (= 0) XR ⁵ , where

X is an oxygen atom, a sulfur atom or a group of the formula NR ⁶ (preferably X = 0) and R ⁵ and R ⁶ independently of one another are a hydrogen atom, an alkyl, Alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aralkyl or heteroaralkyl group or R ⁵ and R ⁶ together form part of a heterocycloalkyl or heteroaryl group are.

R ⁴ is particularly preferably a group of the formula -COOMe.

R ⁴ is more preferably a 4-pyridyl group.

Also preferred are compounds of the general formula (I) in which R ^{1 has} one of the following formulas:

where n is 0, 1 or 2 (preferably 0 or 1; particularly preferably 0).

Compounds of the general formula (I) in which R ^{1 has} the following formula are particularly preferred:

Furthermore, R ¹ is preferably not a group of the formula C (= 0) R ⁷ , where R ^{7 is} a hydrogen atom, an alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, Cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aralkyl or heteroaralkyl group.

Examples of pharmacologically acceptable salts of the compounds of the formula (I) are salts of physiologically acceptable mineral acids such as hydrochloric acid, sulfuric acid and phosphoric acid; or salts of organic acids such as methanesulfonic acid, p-toluenesulfonic acid, lactic acid, acetic acid, trifluoroacetic acid, citric acid, succinic acid, fumaric acid, maleic acid and salicylic acid; or salts of alkali or alkaline earth metal salts such as e.g. B. sodium, potassium, lithium, calcium or magnesium salts, ammonium salts; or salts of organic bases such as e.g. B. methylamine, dimethylamine, triethylamine, piperidine, ethylenediamine, lysine, choline hydroxide, meglumine, morpholine or arginine salts. Compounds of formula (I) can be solvated, in particular hydrated. The hydration can occur during the manufacturing process or as a result of the hygroscopic nature of the initially anhydrous compounds of formula (I). If the compounds of the formula (I) contain asymmetric C atoms, they can be present either as achiral compounds, diastereomer mixtures, mixtures of enantiomers or as optically pure compounds.

A compound or a pharmacological composition of the present invention can be used to inhibit serine proteases (especially tryptase and uPA) and to treat and / or prevent diseases mediated by serine protease activity (especially tryptase and uPA activity). INS special are the described compounds for the treatment of allergic or inflammatory diseases and especially for the treatment of asthma, allergic rhinitis, chronic obstructive pulmonary diseases, emphysema, viral and bacterial lung infections and inflammatory reactions, rheumatoid arthritis, multiple sclerosis, osteoarthritis, dermatological diseases, ulcerative colitis , Conjunctivitis, interstitial cystitis, mastocytosis, psoriasis, conjunctivitis, inflammatory bowel diseases, peptide ulcers, cardiovascular diseases and anaphylaxis of interest. Furthermore, the compounds described here can be used for tumor diseases (both local tumors and tumors in the advanced stage), metastasis formation and cancer.

The pharmaceutical compositions according to the present invention contain at least one compound of formula (I) or a pro-drug thereof as

Active ingredient and optional carriers and / or adjuvants.

The pro-drugs, which are also the subject of the present invention, consist of a compound of the formula (I) and at least one pharmacologically acceptable protective group which is eliminated under physiological conditions, for example an alkoxy, aralkyloxy, acyl or acyloxy group such as an ethoxy, benzyloxy, acetyl or acetyloxy group. A is preferred Hydrogen atom of the amidine replaced by a hydroxy group or alkoxy group.

The therapeutic use of the compounds of formula (I), their pharmacologically acceptable salts or solvates and hydrates as well as formulations and pharmaceutical compositions is also within the scope of the present invention.

The present invention also relates to the use of these active ingredients for the production of medicaments for the prevention and / or treatment of thromboembolytic diseases. In general, compounds of formula (I) are administered using the known and acceptable modes, either individually or in combination with any other therapeutic agent. Such therapeutically useful agents can be administered in one of the following ways: orally, for example as dragées, coated tablets, pills, semi-solids, soft or hard capsules, solutions, emulsions or suspensions; parenterally, for example as an injectable solution; rectally as suppositories; by inhalation, for example as a powder formulation or spray, transdermally or intranasally. For the production of such tablets, pills, semi-solids, coated tablets, dragees and hard gelatin capsules, the therapeutically usable product can be mixed with pharmacologically inert, inorganic or organic drug carrier substances, for example with lactose, sucrose, glucose, gelatin, malt, silica gel, starch or Derivatives thereof, talc, stearic acid or its salts, dry skimmed milk and the like. to In the manufacture of soft capsules, pharmaceutical carriers such as vegetable oils, petroleum, animal or synthetic oils, wax, fat, polyols can be used. For the production of liquid solutions and syrups, drug carriers such as water, alcohols, aqueous salt solution, aqueous dextrose, polyols, glycerin, vegetable oils, petroleum, animal or synthetic oils can be used. For suppositories, pharmaceutical carriers such as vegetable oils, petroleum, animal or synthetic oils, wax, fat and polyols can be used. Compressed gases suitable for this purpose, such as oxygen, nitrogen and carbon dioxide, can be used for aerosol formulations. The pharmaceutically usable agents can also contain additives for preservation, stabilization, emulsifiers, sweeteners, flavorings, salts for changing the osmotic pressure, buffers, coating additives and antioxidants.

For the prevention and / or treatment of those described above

Diseases, the dose of the biologically active compound according to the invention can vary within wide limits and can be adjusted to individual needs. In general, a dose of 0.1 μg to 10 mg / kg body weight per day is suitable, with a preferred dose being 0.5 to 4 mg / kg per day. In suitable cases, the dose can also be below or above the values given above.

The compounds of the general formula according to the invention

(I) can e.g. B. by implementing compounds of Formulas (II) (U. Schöllkopf et al. Liebigs Ann. Chem. 1979, 1444-1446), (III) and (IV) can be produced.

<") (III) (IV)

The reaction is preferably carried out in an alcohol such as. B. ethanol, methanol, isopropanol, butanol or benzyl alcohol (preferably methanol) as a solvent.

The reaction in the presence of a Lewis acid such as. B. boron trifluoride etherate, p-toluenesulfonic acid, ytterbium triflate or indium trichloride.

Again, the reaction is preferably carried out at a temperature of -80 ° C to 100 ° C (again preferably a temperature of -10 ° C to 50 ° C; particularly preferably at room temperature).

The conversion of the methyl ester (for R ⁴ = COOMe) into the different groups of the formulas OR ³ , SR ³ and NR ³ R ⁴ is known to the skilled worker and z. As described in RC Larock, Comprehensive Organic Transformations, VCH Publishers, New York, 1989.

Furthermore, it is possible according to the invention to carry out the reaction on the solid phase, with preference Compound (II) with R ⁴ = COOR ⁵ via R ⁵ on a polymeric support (see e.g. BP Seneci, Solid-Phase Synthesis and Combinatorial Technologies, John Wiley & Sons, New York, 2000; D. Obrecht and JM Villalgordo, Solid -Supported Combinatorial and Parallel Synthesis of Small-Molecular- Weight Compound Libraries, Tetrahedron Organic-Chemistry Series Volume 17, Elsevier Science Ltd, Oxford, 1998; in FZ Dörwald, Organic Synthesis on Solid Phase, Wiley-VCH, Weinheim, 2000) is.

Examples

Assay to determine inhibition of tryptase Tryptase (recombinant human tryptase, Promega) was carried out for 10 min in a buffer (0.1 M Hepes, 0.14 M NaCl, 0.1% PEG

6000, 0.05% Tween 80, pH 7.8 and 200 nM freshly added

Heparin) with various concentrations of the inhibitor

(0.001-1000 μM) incubated. Subsequently, substrate Chromozym PL (Boehringer Mannheim) was added. The

Final enzyme concentration was 3 nM, the

Final concentration of substrate 150μM. The final concentration of DMSO in the assay was 5%. The absorption (405nm) was continuously monitored for 20 min using a Tecan SpectraFluorPlus microtiter plate reader (Tecan,

Crailsheim) followed.

Assay to determine inhibition of uPA uPA (high molecular weight, Calbiochem. No. 672081) was carried out for 10 min in a buffer (0.1 M Hepes, 0.14 M NaCl, 0.1% PEG 6000, 0.05% Tween 80, pH 7.8) with various Concentrations of the inhibitor (0.001-1000 μM) incubated. Subsequently, substrate (Bachern 1-1140) was added. The final concentration of enzyme was 3 nM, the final concentration of substrate 150μM. The fluorescence was monitored continuously for 20 min using a Tecan SpectraFluorPlus microtiter plate reader (Tecan, Crailsheim) (ex: 360nm / era: 465nm).

The enzyme assays were carried out at room temperature in 96 or 384-well microtiter plates. The IC ₅₀ values were calculated using the "GraFit 4" program from Erithacus Software Ltd. (Staines, Middlesex, UK).

General synthesis instructions 1 mmol aldehyde (III) and 1 mmol amine (IV) and 1 mmol

Isocyanide (II) and optionally Lewis acid as

Catalyst are dissolved in 3 ml of methanol and

Room temperature stirred overnight. The solvent is removed in vacuo and the residue is purified by HPLC.

The following compounds were prepared according to the general synthetic procedure. The products were characterized by mass spectroscopy. The tested compounds showed IC ₅₀ values between 0.1 and 10 μM in the tryptase assay described above and between 1 and 50 μM in the uPA assay IC ₅₀ values.

25

Claims

claims

1. Compounds of the general formula (I):

wherein

R ^{1 is} an alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aralkyl or heteroaralkyl group;

R ^{3 is} a hydrogen atom, an alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aralkyl or hetero-aralkyl group and

R ^{4 is} a hydrogen atom, an alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, hetero- is alkylcycloalkyl, aralkyl or heteroaralkyl group;

or a pharmacologically acceptable salt, solvate, hydrate or a pharmacologically acceptable formulation thereof.

Compounds according to claim 1, wherein R ^{2 is} a hydrogen atom.

3. Compounds according to claim 1 or 2, wherein R ^{4 is} a group of the formula -C (= 0) XR ⁵ , where X is an oxygen atom, a sulfur atom or a group of the formula NR ⁶ and R ⁵ and R ⁶ independently of one another is a hydrogen atom, an alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, alkylcycloalkyl, heteroalkylcycloalkyl, aralkyl or heteroaralkyl group or R ⁵ and R ⁶ together form part of a heterocycloalkyl or a heteroaryl group.

4. Compounds according to any one of claims 1 to 3, wherein R ^{4 is} a group of the formula -COOMe.

5. Compounds according to any one of claims 1 to 4, wherein R ^{1 has} one of the following formulas:

where n is 0, 1 or 2.

6. Pharmaceutical compositions containing a compound according to any one of claims 1 to 5.

7. Use of a compound or a pharmaceutical composition according to any one of claims 1 to 6 for the inhibition of serine proteases.

8. Use of a compound or a pharmaceutical composition according to any one of claims 1 to 6 for the treatment and / or prevention of diseases which are mediated by serine proteases.

9. Use of a compound or a pharmaceutical composition according to any one of claims 1 to 6 for the treatment of allergic or inflammatory diseases, tumor diseases and / or cancer.

10th A process for the preparation of a compound according to claim 1, wherein compounds of the formulas (II),

(III) and (IV) are implemented together.

0 ") (III) (IV)